7-cyano steroids



United States Patent Ofiice 3,050,534 Patented Aug. 21, 1962 3,050,534 7-CYANO STEROIDS Albert Bowers and Howard J. Ringold, Mexico City, Mexico, assignors, by mesne assignments, to Syntex Corporation, a corporation of Panama No Drawing. Filed Aug. 13, 1959, Ser. No. 833,387 Claims priority, application Mexico Aug. 13, 1958 37 Claims. ((35. 260-4973) L CHzR CHzOR X & CN

in Which R is a member of the group consisting of hyo drogen, hydroxyl and acyloxy -OR", X is a member of the group consisting of hydrogen or fluorine, Y is an arrangement selected from the group consisting of H OH and =0 Y is either R is a member of the group consisting of hydrogen, the hydroxyl group and acyloxy -OR, the acyl radical in the aforesaid acyloxy groups OR being R" derived from hydrocarbon carboxylic acids having up to about 12 carbon atoms; Z is selected from the group consisting of the single and the double carbon-to-carbon bond, and CN is a member of the group consisting of CN and CN. The R" radical in OR at C17oc can be identical with that at C-2l in the side chain, or-it can be a different acyl radical. The aforesaid hydrocarbon carboxylic acids from which R" is derived, can be of up' to about 12 carbon atoms, saturated or unsaturated, of straight, branched, cyclic or mixed cyclic-aliphatic chain, optionally substituted with functional groups such as hydroxyl. O-acyl (of 1 to about 12 carbon atoms), alkoxy (of 1 to 5 carbon atoms) or halogen (F, Cl or Br). Typical 2l-esters or 17,21-diesters are, among others, the acetate, propionate, butyrate, hemisuccinate, enanthate caproate, benzoate, trimethylacetate, phenoXyacetate, phenylpropionate, cyclopentylpropionate and 3- chloropropionate.

New compounds according to the invention in the pregnane series are represented by the above general Formula B and comprise such 7-cyano pregnane derivatives as the 7a and 7fi-isomers of 7-cyano-progesterone, 7-cyano-17ahydroxyprogesterone, 7 cyano 17oz hydroxy 1 dehydroprogesterone, 7 cyano desoxycorticosterone, 7- cyano-l-dehydro-corticosterone, of the ll-keto, lip-hydroxy, 9u-fluoro-11-keto and 9a-fiuoro-1lfl-hydroxy derivatives of all of the aforementioned compounds, as

' well as of 7-cyano-A -pregnen-17u,21-diol-3,20-dione and 7 cyano A pregnadien l7rx,21 diol 3,20-dione. Our application also comprises the 19 nor derivatives of all of the A -3-ketones mentioned among the above compounds as shown by the general FormulaA. The hydroxyl groups at one or both of the positions C-l7 and C21 may be esterified.

Of the new compounds, the 7-cyano-progesterones and 7-cyano-17a-hydroxyand 17a,21-dihydro'xy hormones (Formula B), particularly under the form of their esters, are potent progestational hormones having antiestrogenic and anti-androgenic activity; the 7-cyano-desoxycorticosterones (7-cyano-cortexones) also exhibit anti-estrogenic and anti-androgenic activity. Furthermore, all of the compounds comprised in our invention, especially those having hydroxyl groups at O-17 and/or 0-21 or an oxygen function at 0-11, are valuable intermediates for the preparation of 7-cyano cortical hormones.

The novel 7-cyano steroidal cortical hormones represented by the above general Formula C of the new compounds according to the invention comprise, more specifically, the and -isomers of 7-cyanocortisone, 7-cyano-cortisol, 7-cyano-prednisone and 7-cyano-prednisolone, which may further have a fluorine atom at 'C-9u and/or may have the hydroxyl group at C-21 under the esterified form. These new compounds which are the object of the present invention, are cortical hormones having a marked anti-inflammatory, glycogenic, thymolytic, eosinopenic, catabolic, anti-androgenic and anti-estrogenic activity.

These several series of new compounds according to our invention are produced by a process using as starting materials steroidal 6-dehydro compounds having the general formulas:

I zR CO CO I? to 1 Alkali or alkaline earth metal cyamide hydroxylated organic solvent (ll aR GO CHzR X Alkali or alkaline earth IA metal eyamide hydroxylated organic solvent CHQR and comprises the introduction of a cyano group at C-7 by refluxing the starting compound with a cyamide ion donator preferably in the form of an alkali metal or alkaline earth metal cyamide in an organic hydroxylated solvent. In the progestational and corticoid series, the

resulting 7-cyano analogs II can be further dehydrogenated to obtain the corresponding A -analogs III.

The 6-dehydro starting compounds having the above general Formulas I and IA are obtained from the corresponding A -3-ketones by applying known methods for dehydrogenation at C6; thus, in the compounds of the IO-methyl series (I) we introduced the double bond at C6,7 by refluxing with chloranil in mixture With xylene (cf. Agnello and Laubach, J. Am. Chem. Soc., 79, 1257 (1957)) or in mixture with ethyl acetate and acetic acid; in the 19-nor-A -3-ketones (IA) we introduced the double bond at C-6,7 by formation of their 3-enol-ethers followed by reaction with hypobromous acid and dehydrobromination of the resulting -6fi-brorno-19-nor-M-3- ketones (cf. Bowers, Casas, Zderic and Djerassi, patent application Serialv No. 806,763, filed on April 16, 1959).

Depending on the nature of the arrangement of substituents at the side chain and at Clc in the starting material, We proceed:

(1) directly with the introduction of a cyano group at C7 by the above described treatment when the side chain is COCH and R is hydrogen or hydroxyl (2) in the case that the arrangement at Cl7 CO. OH OH it is necessary first to protect the l7a,21-diol functions by forming the corresponding 17,20;20,2l-bis-methyleneether, then proceed to introduce the 7-cyano group as described above, and finally reforming the side chain by hydrolyzing the tetrahydropyranyl ether in a conventional manner;

(3) if the arrangement at Cl7 is CO.OH2OH it is necessary first to protect the 17 0:,21-dl0l functions by forming the corresponding l7,20;20,2l'bis-methylenedioxy (BMD) derivatives then introducing the cyano group with C-7 as described under (1), and finally reforming the initial arrangement A at Cl7.

The temporary protection of the dihydroxyacetone side chain can also be achieved by monoketalization of the keto group at C2().

These modes of carrying out the process of producing the new compounds according to the invention are illustrated in more detail in the following Reaction Diagram II, in which X and Y have the same meaning as explained hereinbefore, E designates either hydrogen or the methyl group, and Ac is acetyl.

REACTION DIAGRAM II CHzOAC REACTION DIAGRAM II'Continued" CH3 CH3 I 0 CO A A OH 0H Y Y ,X X E E E o i f lFormaldehyde oHi-o O i 0320 o o I on,

IB IO 0 MN 0N KCN KONl CH -O i CH1 i 0 9 O O 00 /CH t X X E E 0: MN CN 0: NW CN reforming H0 0 0H reforming H C O OH of side chain of side chain $H; (EH3 CHzOH ([JHaOH 00 430 '00 "-OH ---0H Y X X X X E E I E E II II HE 110 0-- MN ON 0: MN ON O W CN 0- AM! CN refluxed with an excess of potassium cyanide in 95% cyanide. After the introduction of the cyano group the ethanol and a mixture of the respective 7a and 7fi-cyanobis-methylenedioxy group was hydrolyzed (for the forma- A -compounds IIA or HA is obtained; the isomers are tion of the 17,20;20,2l-bis-methylenedioxy (BMD) comthen separated by chromatography. For introducing an pounds and their hydrolysis, see Sarrett et al., J. Am. additional double bond at C-1 of the 7-cyano-A -com- Chem. Soc, 80, 1517 (1958)).

pounds of the 10-methy1 series, we reflux the isomers in By the reaction with potassium cyanide we obtained a mixture or after separation with selenium dioxide in mixmixture of the and 7,8-isomers of the 7-eyano-A -3- ture with t-butanol, in the presence of catalytic amounts ketofles, Which Were eparate by chromatography; in the of pyridine and under an atmosphere of nitrogen. BMD compounds the bis-methylenedioxy group was then When starting from A -pregnadiene-17a,21-diol-3,20- 7O hydrolyzed. Thus there were obtained: the 70: and 7f! dione, namely the 6-dehydr0derivatives of cortexolone is m rs of 7-Cyan0-prOgesterone, 7-cyarro-17u-hydroxy- (Reichsteins Compound S) in either th 10- thy1 or progesterone and 7-cyano-cortexone(desoxycorticostethe 19-nor series (IC), we protected the dihydroxyacetone Tone) as W ll as the compounds substituted at C-11 or side chain by formation of its 17,20;20,'21-bis-methy1ene C-11 and C-9, according to the respective starting comdioxy derivative before efiecting the reaction with the 7 pounds, and of 7-cyano-A -pregnene-17a,21-diol-3,20-

the aforesaid free 6-dehydro compounds, there was followed the method described recently by Sarrett et al. (J. Am. Chem. Soc., 80, 1517 (1958)).

The compounds of Formula IC were treated with the cyanide of an alkali or alkali-earth metal in a hydroxylated solvent; for example, they were refluxed with an excess of potassium cyanide in mixture with absolute ethanol for 13 hours. Thus we produced a mixture of the BMD derivatives of the 70: and 7fl-isomers of the 7-cyano analogs of cortisone, hydrocortisone, 9a-fluorocortisone and 9w-fiuoro-hydmeortisone, respectively IC.

By chromatography, for example on washed alumina, we

separated the stereoisomers and obtained 7a-cyano-cortisone, 7a-cyano-hydrocortisone, 7a-cyano-9a-fluoro-cortisome and 7a-cyano-9a-fluoro-hydrocortisone, as Well as REACTION DIAGRAM III the 7fi-isomers of such compounds, respectively, under the form of their BMD derivatives. Upon acid treatment, for example by heating with aqueous formic acid, there was regenerated the dihydroxyacetone side chain and thus we obtained 7a-cyano-cortisone, 7oc-cyano-hydrocortisone, 7a-cyano-9a-fluoro-oortisone and 7a-cyano-9afiuoro-hydrocortisone, as well as the 7 fi-isomers of such compounds, respectively I'Ic.

Usually we first regenerated the dihydroxyacetone side chain from the mixture of the 70: and 75 isomers of 1c and then isolated the stereoisomers by chromatography, to produce the aforementioned compounds of formula Ilc.

While the process of the present invention essentially involved the introduction of the cyano group at 07 into the G-dehydro-derivatives (I), of the compounds which are going to be subjected to such introduction, those resulting A -cyano compounds II which belong to the IO-methyl series, can subsequently be dehydrogenated at 0-1 to produce the respective 7-cyano-1,4-dienes III in the Reaction Diagram I above.

Or the free 17fi-hydroxyl group or the 21-hydroxyl group and, if present, the 17a-hydroxyl group of the above compounds II can be esterified by conventional methods with the radicals of any of the acids specified above, and esterification can then be followed by introduction of a further double bond at C-1. Or the free hydroxyl groups of compounds III may be esterified in the aforesaid manner, as illustrated in the following Reaction Diagram I II:

CH3 llHaoH I O lCO ......RI ---OH X X I H0 0 ON O ON Esterifigatiou i onto-Am I Microbiological Microbiological dehydrogenation dehydrogenation at 0-1 at 0-1 0: NW ON Dehydrogenation with SeOa 311, omon (llHzOAcyl o o 0 o "-R' "-011 ---0B Y Y A l X 1 Saponification 7 III IV V I =O\V CN 0: ON Esterification O M ON 7 I Esterificatlon (l3EzOAcyl C 0 The aforementioned reactions can be modified within wide limits; thus, in the step of introduction of the cyano group, instead of ethanol there can be used another hydroxylated organic solvent such 'as methanol, or aqueous ethanol or aqueous methanol; since this reaction involves the addition of the elements'of hydrogen cyanide to the C-6 double bond, there can be employed any other reagent capable of generating hydrogen cyanide, such as sodium cyanide or the cyanide of an alkaline earth metal.

For introducing as additional double bond into the 7- c3 ano-A -3-ketones of the lO-methyl series, the compounds wererefluxcd with selenium dioxide in mixture with t-butanol, in the presence of catalytic amounts of pyridine and under an atmosphere of nitrogen; in the compounds hydroxylated at 021 the 21-hydroxyl group was previously protected by esterification. .Thus we obtained the 70: and 7,8-cyano derivatives of the aforementioned compounds of the 10-methyl series having an additional double bond at C-l,2; in particular the progesterone 7-cyano derivatives listed above. Among these are the 70: and 7B isomers of 7-cyano-l-dehydro-progesterone, 7- cyano-l-dehydro-l7ot-hydroxyprogesterone, of 21-esters of 7-cya11o-1-del1ydrodesoxycorticosterone, of the corresponding ll-keto(or f3-hydroxy) and 11-keto(or {i-hydroxy-9a-fluoro-compounds, as well as the 70: and 7,8 isomers of 21-esters, preferably the acetate, of A pregnadiene-17a,21-dione.

The ester group at C-21 of all of the 21-esters of the 7- cyano compounds was hydrolyzed and the hydroxyl groups which might be present at one or both of the positions C-17 and C-21 were reesterified; for eifec ting these operations we employed conventional methods which for clarity are described in some examples.

The Zl-hydroxyl group of the compounds of Formula He was esterified by reaction with an excess of the anhydride of a carboxylic acid of up to 12 carbon atoms, in pyridine solution, to produce their 21-esters IV.

In another mode of carrying out the process for producing the A steroidal 7-cyano derivatives according tothe invention, the respective 7-cyano compound of the 10- methyl series (in the case of the alcohols preferably under the free form) for instance, the 21-hydroxy compounds Ilc were incubated with cultures of Corynebacterium simplex ATCC 6946 or of Septomia ATCC 6737-, although there can be used other species of microorganisms. For this microbiological dehydrogenation one can also start from a 21-ester of the steroid (IV) obtained as mentioned above, since these methods involve the simultaneous hydrolysis of the ester group at C-21; in any case the resulting product is the respective 21-hydroxy-7- cyano-A -compound III, that is, for example, one of the following important compounds: 7u-cyano-prednisone, 7a-cyano-9a-fiuoro-prednisone, 7a-cyano-9a-fluoroprednisolone, as well as the 7fi-isomers of such compounds, respectively. By the aforementioned reaction with the anhydride of a carboxylic acid, in pyridine solution, there were obtained the corresponding 2l-esters V.

We also obtained the esters of Formula V by dehydrogenation of the 2l-esters of the 7-cyano-A -compounds IV by refluxing with selenium dioxide, preferably in mixture with t-butanol, in the'presence of catalytic amounts of pyridine and under an atmosphere of nitrogen.

By alkali treatment, for example with dilute methanolic potassium hydroxide or sodium methoxide solution at low temperature and under an atmosphere of nitrogen, there can be hydrolyzed the ester groups of IV and V; the 21-hydroxyl group can be re-esterified by the aforementioned method. 7

For the esterification at C-2l we employed the anhydrides of carboxylic acids of up to 12 carbon atoms, saturated or unsaturated, of straight, branched, cyclic or mixed cyclic-aliphatic chain, which acid may be substituted with groups such as methoxy, halogen or other groups; among other such esters we prepared the 21- acetates, propionates, t-butyrates, hemisuccinates, enanthates, caproates, benzoates, .trimethylacetates, phenoxyacetates, cyclopentylpropionates, phenylpropionates and ,B-chloropropionates.

The cyano group at C-7 of the present novelcornpounds can be transformed into other groups For eX- ample, it can be converted into a carboxyl group, which in turn can produce a methyl ester which can be reduced to the hydroxymethylene group. V r

The starting materials I and IA are well known in the steroid art;'of those less frequently used, the Qua-fluoroanalogs of ll-keto-progesterone, l7a-l'1YdIOXY-1l-lifit0- progesterone, llfl-hydroxyprogesterone and 11 3, 17mdihydroxy-progesterone were described by Fried et al. in J. Am. Chem. Soc. 77, page 1068 (1955). Of the norcompounds 19'nor-cortexone 19-1101 Desoxycorticosterone) was described by Sandoval et al. in J. Am. Chem. Soc. 77, page 148-(1955), 1IB-hydroxy-l9-nor-progesterone and. 1l-keto-l9 nor-progesterone were. described by Bowers et al. in Tetrahedron (l958), vol. 2, page 165, and 17a-hydroxy-l9-nor-progesterone by Zafl'aroni, et al. in I. Am. Chem. Soc. 76, page 6210 (1950).. 9a-fiuoro- 11 3-hydroxy-19-nor-progesterone and 9oc-fll10rO-1 1-keto- 19-nor-progesterone are described in patent application SerialNo. 760,053,filed September 10, 1958, now

A mixture of 5 g. of 9 t-fiuoro-cortisone 2lace tate prepared as described byFried et al. in US. Patent 2,852,511 and by the same authors in Recent Progress in Hormone Research vol. XI (1955) p. 168 et seq., 3 g. of chloranil and 200.cc..of xylene was refluxed for. 12 hours, cooled and diluted with ether. Thesolution was washed with water, 5% sodium carbonate solution and again withwater to neutral, dried over anhydrousysodi'um sulfate and evaporated to dryness, finallyunder, vacuum. By chromatography of the residue on ethyl acetate washed alumina there was obtained 9u-fluoro-A -pregnadiene- 17a,21-diol-3,11,20-trione 2l-acetate, that is, 9 -fluoro-6- dehydro-cortisone Zl-acetate. i

A suspension of 3. g. of the above compound in 30 cc. of absolute methanolwas treated with a 3Q cc. of a so: lution of sodium methoxide in methanolprepared from mg. of sodium metal, and the mixture was stirredfor 1 hour at 0* C. under an atmosphere of nitrogen. The mixture was then poured into aqueous saturated sodium chloride solution containing 0.5 cc. of acetic acid and the precipitate was collected by filtration, washed with water, dried and recrystallized from acetone-hexane, There was thus.obtained thewfree 9 -fiuoro-6-dehydrocortisone. i l t p V 1 Preparation 11 By following the method of the previous preparation, there was introduced an additional double bond between G6 and. C-7 'of" 9a-fiuoro-cortisol"21*acetate and t the acetate group was then hydrolyzed yielding the free-9afluoro-6-dehydro-cortisol (cortisol=hydroctis0ne) l Preparation III f I V By the reaction with sodiumwmethoxide; described in Preparation I, the known 21acetates o'f 6-dehydro-cortisome and 6-dehydrohydrocortisone were converted into the respective free alcohol. t

Pfeparation IV washed alumina yielded 6-dehydro-l7a-hydroxy-progesterone.

Preparation V Applying the procedure of the preceding preparation to 17a-hydroxy-l9-nor-progesterone produced as described by Zaffaroni et al. in J. Am. Chem. Soc. 76, page 6210 (1950), there was obtained l7u-hydroxy-6-dehydro-19- nor-progesterone.

Preparation VI Applying the procedure of the preceding Preparation IV to 9a-fluoro-1l-keto-progesterone produced as described by Fried et a]. in I. Am. Chem. Soc. 77, page 1068 (1955), there was obtained 6-dehydro-9a-fluoro-1l-keto-progesterone.

Preparation VII By applying the procedure described by Sondheimer et al. in I. Am. Chem. Soc. 75, page 1932 et seq., 1953, there is produced 6-dehydro-oortexone 21-acetate which is saponified in a conventional manner to obtain the free 6-dehydro-cortexone.

EXAMPLE I A mixture of 2.5 g. of G-dehydro-cortisol (prepared as described by Hershberg and co-workers in I. Am. Chem. Soc. 79, page 502 (1957)), 100 cc. of chloroform and 25 cc. of a 37% aqueous solution of formaldehyde was treated with 25 cc. of concentrated hydrochloric acid and :stirred at room temperature for 48 hours. The chloroform layer was separated, washed with aqueous sodium carbonate solution and then with water to neutral, dried over anhydrous sodium sulfate and the chloroform was evaporated. Recrystallization of the residue from acetone-hexane yielded l7,20 ;20,2l-bis-methylene-dioxy A -pregnadiene-3,1l-dione, that is, the bismethylenedioxy derivative of hydrocortisone.

A mixture of 5 g. of 17,20;20,21-bis-methylenedioxy- A -pregnadien-l1 6-ol-3-one (BMD derivative of 6-dehydro-hydrocortisone), g. of potassium cyanide and 250 cc. of absolute ethanol was refluxed for 2 /2 hours, cooled and diluted with water. The product was extracted with ethyl acetate, washed with water, dried over anhydrous sodium sulfate and evaporated to dryness. There was thus obtained a mixture of the 7uand 7.,8-isomers of 7-cyano-l7,20;20,21 bis methylenedioxy-A pregnen- 11,B-ol-3-one in crude form, which can be separated into its components by chromatography.

The crude bis-methylenedioxy (BMD) derivative of the above 7-cyano-hydrocortisone was mixed with 500 cc. of

60% formic acid and heated on the steam bath for minutes, cooled and diluted with aqueous saturated sodium chloride solution; the precipitate was collected, washed with a little cold water, dried and recrystallized from acetone-hexane. There was thus obtained a mixture of 7acyano-hydrocortisone and 7,6-cyano-hydrocortisone. The stereoisomers were separated by chromatography on neutral alumina.

In each of Erlenmeyer flasks of 125 cc. capacity there was placed 30 cc. of an aqueous medium of 1% yeast extract. The contents of each flask was inoculated with a culture of Corynebacterium simplex ATCC 6946, obtained by incubating an aqueous medium of 1% yeast extract with a suspension of such microorganism for 24 hours. The medium thus inoculated was stirred for 24 hours at 28 C. There was thus obtained the culture of Corynebacterium simplex which was used for the subsequent incubation with the steroid.

To each flask there was added 1 cc. of a 1% solution of 7a-cyano-hydrocortisone in ethanol, which solution had been prepared just before its use and in recently distilled ethanol. The mixtures were then incubated at 26 C. for 24 hours with continuous stirring; in other experiments the incubation period was extended up to 72 hours.

The contents of the flasks were combined and divided into 3 portions; each portion was extracted 5 times with 5 00 cc. portions of methylene chloride. The extracts were combined, washed with waterdried over anhydrous sodium sulfate and the methylene chloride was evaporated. The residue was purified by chromatography on neutral alumina. The fractions eluted with mixtures of benzene and ether were crystallized from ethyl acetate, thus yielding 7 a-cyano-prednisolane.

EXAMPLE 11 Free 9a-fluoro-6-dehydro-cortisone obtained as de scribed in Preparation I was treated in the same manner as described for the corresponding cortisol derivative in Example I and was first transformed into its bis-methylenedioxy derivative, to produce the bis-methylenedioxy derivative of 9oc-fl1lOI'O-6-dChYdrO-COIli80l16, then to 7ot-cyanocortisone, and finally to 7a-cyano prednisone.

EXAMPLE III By following the microbiological dehydrogenation method described in Example I, 7j3-cyano-hydro-cortisone was incubated to produce 7fi-cyan0-prednisolone.

EXAMPLE IV A culture of Sepzomixa afifnis ATCC 67 37 was prepared by inoculating an aqueous medium of 2% of peptone and 5% of corn syrup with a growing vegetating culture of such fungus in the same medium. It was then incubated for 24 hours at a temperature around 28 C. under continuous stirring.

To each lt. of the Streptomyxa culture thus obtained there was added 30 cc. of a 1% solution of 7a-cyano-hydrocortisone in recently distilled ethanol and prepared just before its use. The mixture was stirred for 24 hours with aeration at a temperature around 28 C.; in other experiments the incubation period was extended up to 72 hours.

The product was then extracted several times with methylene chloride and the extract was washed with water, dried over anhydrous sodium sulfate and concentrated to a small volume under reduced pressure. The residue was subjected to chromatography on neutral alumina, eluting with mixtures of benzene and ether. By recrystallization from ethyl acetate of the fractions thus obtained, there was produced 7a-cyano-prednisolone, identical with the one obtained in accordance with the method of Example I.

EXAMPLE V By substituting in the method of the previous example the 7a-cyano-hydrocortisone by 7,8-cyano-hydrocortisone there was obtained 7fl-cyano prednisolone, identical with that obtained in accordance with Example III.

EXAMPLE VI A solution of 3 g. of 7a-cyano-hydrocortisone in 15 cc. of pyridine was treated with 3 cc. of acetic anhydride and the mixture was allowed to react overnight at room temperature. It was then poured into water, heated for half an hour on the steam bath, cooled and the precipitate was collected, washed with water, dried and recrystallized from acetone-hexane. There was thus obtained 7ot-cyanohydrocortisone 21-acetate.

A mixture of 3 g. of the above compound, cc. of t-butanol, 1.2 g. of selenium dioxide and 1 cc. of pyridine was refluxed under an atmosphere of nitrogen for 48 hours, filtered through celite, washing the filter with a little hot ethyl acetate and the filtrate and washings were combined and evaporated to dryness under reduced pressure. The residue was dissolved in acetone, refluxed with charcoal for 1 hour, filtered and the acetone was evaporated. The residue was purified by chromatography on neutral alumina. By recrystallization of the solid eluates from acetone-hexane there was obtained 7ot-cyano-prednisolone ZI-acetate.

A suspension of 1 g. of the above compound in 10 cc.

13 of absolute methanol was cooled to 0., treated with a solution of sodium methoxide prepared by dissolving :60 mg. of sodium metal in cc. of absolute methanol and the mixture was stirred at 0 C. for 1 hour; it was then poured into 100 cc. of aqueous saturated sodium chloride solution containing a few drops of acetic'acid and the precipitate was collected, washed with water, dried and recrystallized from acetone-hexane. There was thus obtained 7a-cyano-prednisolone, identical with the one obtained in accordance with Examples I and IV.

EXAMPLE VII By following the method of Example I, the BMD derivative of 6-dehydro-cortisone was treated with potassium cyanide to produce a mixture of the 7-stereoisomers of the BMD derivatives of 7-cyano-cortisone; then the dihydroxyaeetone side chain was regenerated and the stereoisomers were separated by chromatography. By following the procedures described in the previous examples, there were then prepared the 7,B-cyano-prednisone and its 21- acetate, respectively.

EXAMPLE VIII By substituting in. the method of Example I for the BMD derivative of 6-dehydro-hydrocortisone of the BMD derivative of Qa-fluoro-6-dehydro-cortiso1 obtained as described in Preparation II, there were prepared: 7a-cyano- 9a-fluoro-cortisol, 7cc-CY3I10-9zx-flllO*IO-pl6ClI1iSOl0Il6, their 7y3-isomers and the 2l-acetates of the aforementioned 7- cyano-compounds.

EXAMPLE IX By using in the method of Example I 9a-fluoro 6-dehydro-cortisone obtained as described in Preparation I as starting material, there were prepared: 7oc-CY3I10-9ccfluoro-cortisone, 7a-cyano-9a-fiuoroprednisone, their 7 8- epirners, and the 21-acetate of the aforementioned 7- cyano compounds.

By following themethod of acetylation described in Example VI, there was esterified the 2l-hydroxyl group of the 7m and 7 3-isorners of 7-cyano-hydrocortisone, of 7- cyano-cortisone, 7-cyano-prednislone and of 7-cyano-prednisone, as well as of the 9a-fluoro-analogs of such compounds, but using instead of acetic anhydride the anhydride of any other carboxylic acid of up to 12 carbon atoms. In the case of polycarboxylic acids there was used a larger quantity of the anhydride and the reaction time was extended to 48 hours.

In following this procedure of Example VI the final products listed in Table I below can be obtained from the indicated starting materials:

EXAMPLES XVII-XXIII By dehydrogenation of the 21-esters of the 7-cyano-A compounds, by refluxing with selenium dioxide as described in Example VI, there were obtained the corresponding 21-esters of the 7- cyano-A -compounds, as listed below in Table II.

TABLE II Ex. No. 7-oyau0-N-eompound 7-cyano-A -c0mpound 5 XVII. 7o: cyano cortisol 21 -pr0- 7a cyano prednisolone 21- pionate. propiouate. XVIIL... 7 8 cyano cortisone 21- 7,8 cyano prednisone 21- butyrate. butyrate. XIX 7,8 cyano 9oz fiuoro cor- 7 cyano 9o: fluoro predtisol-2l-enanthate. nisolone-Zl-nantliate. XX 7oz cyano 9a fiuoro corti 7a cyano 9oz fluoro pred- 1 G sone-Zlhemisuecinatc. nisone-El-hernisuccmate.

XXI 7,8 cyano cortisol 21 ben- 75 cyano prednxsolone 21 zoate. benzoate. XXII. 7oz cyano cortisone 21- 7a cyano prednisone 21- caproate. caproate. XXIII 7,9 cyano cortisone 21- 75 cyano predmsone 21- eyclopentyhpropionate. cyclopentyl-propionate.

EXAMPLE XXIV My reaction of 7oc-CYfiI10-PI6ClIliSOlOI16-2I-PIOPIOIIBIC, obtained according to Example XVII, with sodium methoxide, in accordance with Example VI, there was obtained the free 7a-cyano-prednisolone.

EXAMPLE XXV By incubation of a mixture of the 21-butyrates of 7aand 7fi-cyano-cortisones, by following the procedure described in Example IV, there was obtained a mixture of the 70: and 7B-isomers of respective 7-cyano-prednisones having the hydroxyl group at C-21 under the free form;

EXAMPLE XXVI EXAMPLE XXVII By applying the method described in Example XXVI to 5 g. of 17whydroxy 6-dehydro-progesterone obtained as described in Preparation IV there were produced 71x-CY21I10- l7a-hydroxy-progesterone and 7B cyano Uni-hydroxyprogesterone.

EXAMPLE XXVIII By applying the method of Example XXVI to 5 g. of l7a-hydroxy-19-nor-6-dehydro-progesterone produced according to Preparation V there were obtained 7a-cyano- 17oc-l1YdIOXY l9 nor-progesterone and 7B-cyano-17ahydroxy-19-nor-progesterone.

EXAMPLE XXIX By applying the method of XXVI to 5 g. of 11-keto-9afluoro-6 dehydro-progesterone produced according to Preparation VI there were obtained 7a-cyano-11-keto-9ufluoro-progesterone and 7,8-cyano 11' l(O-9ocflL1OIO- progesterone.

EXAMPLE XXX A mixture of 5 g. of 6-dehydr0-cortexone (cortexonedesoxycorticosterone) obtained as described in Preparation VII, 60 cc. of dry benzene, 6 cc. of dihydropyrane and 120 mg. of p-toluenesulfonic acid was kept at room temperature for 20 hours and then poured into aqueous saturated sodium bicarbonate solution; the organic phase was separated, washed with water, dried over anhydrous sodium sulfate and evaporated to dryness, thus producing the 21-(2'-tetrahydropyranyl)-ether of 6-dehydro-cortexone.

In accordance with the method described in Example XXVI, 5 g. of the above compound was treated with po- 75 tassium cyanide to form the 2l-(2'-tetrahydropyranyl)- The product I dryness.

aoeoeea 15 ether of 7u-cyano-cortexone and of 7/3-cyano-cortexone, respectively.

The ether group of the above compounds was hydrolyzed in the usual manner by treatment in acetone solution with small amounts of concentrated hydrochloric acid; the mixture was then diluted with water and the product was extracted with ethyl acetate and purified by recrystallization from acetone-hexane.

EXAMPLE XXXI A mixture of g. of 6-dehydro-cortexolone (M -pregnadiene-l7a,21-diol-3,20-dione), prepared by conventional saponification of the 21-acetate thereof obtained as described by Sondheimer et al. in J. Am. Chem. Soc. 75, pages 5932 et seq. (1953), 150 cc. of chloroform, 50 cc. of 37% aqueous formaldehyde solution and 50 cc. of concentrated hydrochloric acid was stirred at room temperature for 48 hours. The chloroform layer was separated, washed with 5% aqueous sodium carbonate solution and water, dried over anhydrous sodium sulfate and evaporated to dryness. Recrystallization of the residue from acetone-hexane aflorded 17,20;20,2l-bis-methylenedioxy-A -pregnadien-3-one.

A solution of 3 g. of the above compound in 95% ethanol was treated with potassium cyanide and the product was worked up in accordance with the method described in Example I. The crude product was mixed with 300 cc. of 60% formic acid and heated on the steam bath for 20 minutes, cooled, diluted with aqueous saturated sodium chloride solution and the precipitate formed was collected by filtration, Washed with Water and dried. By chromatography on neutral alumina there were obtained 7a-cyano-A -pregnene-17a,2l-diol-3,2O-dione and 7fl-cyano-M-pregnene-l7a,21-diol-3,20-dione.

EXAMPLE XXXII A mixture of 2 g. of 7-cyano-progesterone (Example I), prepared as described in Example XXVI, 100 cc. of t-butanol, 250 mg. of selenium dioxide and a few drops of pyridine was refluxed under an atmosphere of nitrogen for 48 hours, filtered through celite and evaporated to The residue was dissolved in acetone, treated with decolorizing charcoal, refluxed for 1 hour, filtered and again evaporated to dryness. By chromatography of the residue on neutral alumina there was obtained 7OL-Cyano-l-dehydro-progesterone.

EXAMPLE XXXIII By following the procedure of the previous example, 3 g. of 7fl-cyano-17a-hydroxy-progesterone (Example 2) was converted into 7fl-cyano-17a-hydroxy-1-dehydroprogesterone.

EXAMPLE XXXIV A solution of 3 g. of 7ot-cyano-cortexone, obtained as described in Example XXX, in cc. of pyridine and 3 cc. of acetic anhydride was kept for 4 hours at room temperature, poured into Water, heated for half an hour on the steam bath, cooled and the precipitate was collected by filtration, washed with water, dried and recrystallized from acetone-hexane. There was thus obtained 7u-cyano-cortexone 21-acetate.

2 g. of the above compound was refluxed with selenium dioxide, as described in Example 7, to produce 7a-cyano l-dehydro-cortexone 21-acetate.

A mixture of l g. of the above compound and 50 cc. of a 1% methanolic solution of potassium hydroxide was kept at 0 C. under an atmosphere of nitrogen for 1 hour; it was then poured into 200 cc. of cold sodium chloride solution containing 1 cc. of acetic acid and the precipitate was collected, washed with water, dried and recrystallized from acetone-hexane, thus yielding the free 7ot-cyano-1- dehydro-cortexone.

A mixture of 500 mg. of the above compound, 5 cc.

155 of pyridine and 2 cc. of benzoic anhydride was kept overnight .at room temperature; after pouring into cc. of water the mixture was heated for 2 hours on the steam bath, cooled and the precipitate was collected, washed with water, dried and recrystallized from acetone-hexane, thus furnishing 7a-cyano-1-dehydro-cortexone 21-benzoate.

EXAMPLE XXXV A mixture of 2 'g. of 7u-cyano-cortexolone (ht-cyano- A pregnene 17a,21 diol 3,20 dione) prepared as described in Example XXXI, 10 cc. of pyridine and 4 g. of cyclopentylpropionic anhydride was kept for 48 hours at room temperature, poured into water, heated for 2 hours on the steam bath, cooled and the precipitate was collected, washed with water, dried and recrystallized from acetone-hexane. There was thus obtained 7a-cyano-cortexolone 2l-cyclopentylpropionate.

By reaction of 1 g. of the above compound with selenium dioxide, in accordance with the procedure described in Example XXXII, there was obtained the corresponding 21-ester of 7ot-cyano-l-dehydro-cortexolone, i.e., 7a cyano A pregnadiene 17ot,21 diol 3,20 dione 21-cyclopentylpropionate.

A solution of 500 mg. of the above compound and 500 mg. of p-toluenesulfonic acid in a mixture of 15 cc.

of acetic acid and 3 cc. of acetic anhydride was kept overnight under an atmosphere of nitrogen; after pouring into water the mixture was heated for half an hour on the steam bath, cooled and the precipitate was collected, Washed with Water, dried and recrystallized from acetonehexane. There was thus obtained 7nt-cyano-A -pregnadiene l7 x,21 diol 3,20 dione 17 acetate 21- cyclopentylpropionate.

EXAMPLE XXXVI A mixture of 1 g. of 7ot-cyano-17m-hydroxy-19-norprogesterone prepared as described in Example XXVIII, 100 cc. of anhydrous benzene, 3 g. of caproic anhydride and 200 mg. of p-toluenesulfonic acid monohydrate was kept at room temperature for 3 days and then diluted with Water. The benzene layer was separated, washed with 5% sodium carbonate solution and water, dried over anhydrous sodium sulfate and the benzene was evaporated. By chromatography of the residue on neutral alumina there was obtained 7ot-cyano-17ot-hydroxy-19-norprogesterone caproate.

We claim:

1. A new steroid compound having the general formula:

in which R and R' are members of the group consisting of hydrogen, hydroxyl, and acyloxy OR", R is an acyl radical derived from a hydrocarbon carboxylic acid having up to about 12 carbon atoms, X is a member of the group consisting of hydrogen and fluorine, Y is an arrangement selected from the group consisting of H OH and =0 H H and CN represents a cyano radical selected from the group consisting of 7ot-cyano and 7,B-cyano.

2. A new steroid compound having the general formula:

in which R and R are members of the group consisting of hydrogen, hydroxyl, and acyloxy OR, R" is an acyl radical derived from a hydrocarbon carboxylic acid having up to about 12 carbon atoms, X is a member of the group consisting of hydrogen and fluorine, Y is an arrangement selected from the group consisting of and and WCN represents a cyano radical selected from the group consisting of 7 a-cyano and 7 fl-cyano, and in which Z designates a C-l-to-C-Z linkage selected from the group consisting of C-C and C=C.

3. 7-cyano cortisone.

4. 7-cyano cortisol.

5. 7-cyano prednisone.

6. 7-cyano prednisolone.

7. A steroid as described in claim 2, wherein X is fluorine and Y is =0.

8. A steroid as described in claim 2, wherein X is fluorine and Y is 9. A steroid as described in claim 7, wherein Z is a double bond.

10. A steroid as described in claim 8, wherein Z is a double bond.

11. 7-cyano progesterone. 12. 7-cyano-17a-hydroxy-progesterone. 13. 7-cyano-cortexone. l4. 7-cyano-cortexolone. 15. 7-cyano-19-nor-progesterone. 16. 7-cyano-19-nor-17a-hydroxy-progesterone.

7-cyano-19-nor-cortexone. 7-cyano-19-nor-cortexolone. 7-cyano-l-dehydro-progesterone. 7-cyano-17u-hydroxy1-dehydro-progesterone. 7-cyano-l-dehydro-cortexone. 7-cyano-l-dehydro-cortexolone. In a process for producing 7-cyano steroids, the step of refluxing a starting 6-dehydro-steroid selected from the group consisting of compounds having the general formula CHzR H on and =0 and E is selected from the group consisting of hydrogen and methyl, and their l7,20;20,21-bis-methylenedioxy derivatives with an excess of a cyanide salt selected from the group consisting of alkali metal cyanides and alkaline earth metal cyanides in solution in a hydroxylated organic solvent.

24. The process step described in claim 23, in which the cyanide salt is potassium cyanide dissolved in ethanol.

25. A 21-mono hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano cortisone.

26. A 21-mono hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano cortisol.

27. A 21-mono hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano prednisone.

28. A 2.1-mono hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano prednisolone.

29. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-17a-hydroXy-progesterone.

30. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-cortexone.

31. A hydrocarbon carboxylic acid ester of carbon atoms of 7-cyano-cortexolone.

32. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-l9-nor-17a-hydroxy-progesterone.

33. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-19-nor-cortexone.

34. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-19-nor-cortexolone.

35. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-17u-hydroxy-l-dehydro-progesterone.

36. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-l-dehydro-cortexone.

37. A hydrocarbon carboxylic acid ester of up to 12 carbon atoms of 7-cyano-l-dehydro-cortexolone.

up to 12 References Cited in the file of this patent UNITED STATES PATENTS 2,841,531 Wettstein et a1. July 1, 1958 2,888,457 Beyler et a1. May 26, 1959 2,897,218 Sebech et a1 July 28, 1959 2,907,694 Agnello et a1. Oct. 6, 1959 

1. A NEW STEROID COMPOUND HAVING THE GENERAL FOARMULA: 